In order to treat successfully a disease caused by a bacterium, the rapid and accurate detection and identification of the disease-causing bacterium is required. The detection and identification have traditionally been accomplished by pure culture isolation and identification procedures that make use of knowledge of specimen source, growth requirements, visible (colony) growth features, microscopic morphology, staining reactions, and biochemical characteristics.
A number of different bacterial species can cause meningitis when present in the CSF. The species most frequently causing meningitis include: Escherichia coli and other enteric bacteria, Haemophilus influenzae, Neisseria meningitidis, Streptococcus pneumoniae, Streptococcus agalactiae, and Listeria monocytogenes.
Conventional methods of detection and identification of bacteria in cerebrospinal fluid include the Gram stain, latex agglutination and other antibody-based tests, and culture. The Gram stain and antibody-based tests are rapid (&lt;1 hour), but of low sensitivity (requiring at least 10.sup.4 colony forming units [CFU]bacteria per ml). Culture methods, while sensitive to approximately 2 CFU per ml, require overnight incubation.
A number of scientific publications relating to this invention exist. For example, the polymerase chain reaction has been used to detect individual species of bacteria causing meningitis: Kuritza and Oehler, May, 1991, Abstracts of the General Meeting of the ASM page 84; Deneer and Boychuk, 1991, Applied Environmental Microbiology 57: 606-609; and Kristiansen et al., 1991, Lancet 337: 1568-1569.
In addition, some of the nucleotide sequence data used herein is available in Genbank. The method of reverse dot-blot detection has been described by Saiki et al., 1989. The use of uracil-N-glycosylase has been described by Longo et al., 1990, Gene 93: 125-128.
A method of detecting bacteria in Cerebrospinal Fluid ("CSF") which is both sensitive and rapid would represent a great improvement over current methods of detection. The present invention meets these needs.